[Code of Federal Regulations]
[Title 9, Volume 1]
[Revised as of January 1, 2004]
From the U.S. Government Printing Office via GPO Access
[CITE: 9CFR113.47]

[Page 614-615]
 
                  TITLE 9--ANIMALS AND ANIMAL PRODUCTS
 
  CHAPTER I--ANIMAL AND PLANT HEALTH INSPECTION SERVICE, DEPARTMENT OF 
                               AGRICULTURE
 
PART 113_STANDARD REQUIREMENTS--Table of Contents
 
Sec.  113.47  Detection of extraneous viruses by the fluorescent 
antibody technique.

    The test for detection of extraneous viruses by the fluorescent 
antibody technique provided in this section shall be conducted when 
prescribed in an applicable Standard Requirement or in a filed Outline 
of Production for a product.
    (a) Monolayer cultures of cells (monolayers), at least 7 days after 
the last subculturing, shall be processed and stained with the 
appropriate antiviral fluorochrome-conjugated antibody as specified in 
paragraph (b) of this section.
    (1) Three groups of one or more monolayers shall be required for 
each specific virus prescribed in paragraph (b) of this section.
    (i) At the time of the last subculturing, one group of test 
monolayers shall be inoculated with approximately 100-300 
FAID50 of the specific virus being tested for as positive 
controls.
    (ii) One group of monolayers shall be the ``material under test.''
    (iii) One group of monolayers, that are of the same type of cells as 
the test monolayers and that have been tested as prescribed in 
Sec. Sec.  113.51 or 113.52 (whichever is applicable), shall be prepared 
as negative controls.
    (2) Each group of monolayers shall have a total area of at least 6 
cm\2\.
    (3) Positive control monolayers may be fixed (processed so as to 
arrest growth and assure attachment of the monolayer to the surface of 
the vessel in which they are grown) before 7 days after subculturing if 
fluorescence is enhanced by doing so, Provided, That a monolayer of the 
material under test is also fixed at the same time as the positive 
control and a monolayer of the material under test is also fixed at 
least seven days after subculturing. Monolayers that are fixed before 7 
days after subculturing shall be stained at the same time as the test 
monolayers and negative controls fixed at least 7 days after 
subculturing.

[[Page 615]]

    (b) The antiviral fluorochrome-conjugated antibodies to be used 
shall depend on the type of cells required to be tested for extraneous 
viruses as specified in an applicable Standard Requirement or in a filed 
Outline of Production. Antiviral fluorochrome-conjugated antibodies 
specific for the extraneous viruses shall be applied to each respective 
type of cell in accordance with the following list. Under certain 
circumstances, additional tests may need to be conducted, as determined 
by the Administrator. When a specific antiviral fluorochrome-conjugated 
antibody is used in testing for the listed extraneous viruses specified 
in more than one cell type, it need only be applied to the most 
susceptible cell type.
    (1) All cells shall be tested for:
    (i) Bovine virus diarrhea virus;
    (ii) Reovirus; and
    (iii) Rabies virus.
    (2) Bovine, caprine, and ovine cells shall, in addition, be tested 
for:
    (i) Bluetongue virus;
    (ii) Bovine adenoviruses;
    (iii) Bovine parvovirus; and
    (iv) Bovine respiratory syncytial virus.
    (3) Canine cells shall, in addition, be tested for:
    (i) Canine coronavirus;
    (ii) Canine distemper virus; and
    (iii) Canine parvovirus.
    (4) Equine cells shall, in addition, be tested for:
    (i) Equine herpesvirus; and
    (ii) Equine viral arteritis virus.
    (5) Feline cells shall, in addition, be tested for:
    (i) Feline infectious peritonitis virus; and
    (ii) Feline panleukopenia virus.
    (6) Porcine cells shall, in addition, be tested for:
    (i) Porcine adenovirus;
    (ii) Porcine parvovirus;
    (iii) transmissible gastroenteritis virus; and
    (iv) Porcine hemagglutinating encephalitis virus.
    (7) Firms that do not have rabies virus on premises either for 
research or production purposes are exempt from having to produce 
positive rabies virus control monolayers. Fixed positive rabies virus 
control monolayers will be provided by the National Veterinary Services 
Laboratories.
    (c) After staining, each group of monolayers shall be examined for 
the presence of specific fluorescence attributable to the presence of 
extraneous viruses.
    (1) If the material under test shows any evidence of specific viral 
fluorescence, it is unsatisfactory and may not be used; Provided, That, 
if specific fluorescence attributable to the virus being tested for is 
absent in the positive control monolayers, the test is inconclusive and 
may be repeated.
    (2) If the fluorescence of the monolayers inoculated with the 
specific virus as positive controls is equivocal, or if the negative 
monolayers show equivocal fluorescence indicating possible viral 
contamination, or both, the test shall be declared inconclusive, and may 
be repeated; Provided, That, if the test is not repeated, the material 
under test shall be regarded as unsatisfactory for use in the production 
of biologics.

[60 FR 24548, May 9, 1995]

                         Ingredient Requirements